triple negative breast cancer cell lines hs578t  (ATCC)

Journal: Cancers

Article Title: Zebrafish Xenografts Unveil Sensitivity to Olaparib beyond BRCA Status

doi: 10.3390/cancers12071769

Figure Lengend Snippet: Zebrafish xenografts of triple negative breast cancer (TNBC) reveal different responses to olaparib and ionizing radiation (IR) independently of BRCA status. TNBC cell lines (Hs578T BRCAwt , MDA-MB-469 BRCAwt and Hcc1937 BRCA1mut ) were fluorescently labeled with CM-DiI (not shown) and injected into the perivitelline space (PVS) of 2dpf zebrafish embryos. At 24 hpi, zebrafish xenografts were screened and randomly distributed into the different experimental conditions: control, olaparib, IR, IR + olaparib. Xenografts were treated for 4 consecutive days and fixed at 5dpi. Representative confocal images of xenografts labelled with anti-human-leukocyte antigen-major histocompatibility complex (HLA-MHC)-class I (in red) ( A,E,I ) and anti-activated caspase 3 (in green) ( A’ – L ). Nuclei were stained with DAPI (in blue) ( A – L ). Mitotic index ( M) , nuclear area size ( N ), cell death-activated caspase3 ( O ) and average tumor size (number of human DAPI cells) ( P ) were analyzed by confocal microscopy and quantified. The % of activated caspase 3 cells and tumor size were normalized to respective controls to compare between different xenografts in different conditions. All images are anterior to the left, posterior to right, dorsal up, and ventral down (as depicted in the scheme on top left). The dashed line delineates the tumor area. Scale bar: 50 µm. Results are from three independent experiments and expressed as mean ± SEM, each dot represents one xenograft. The total number of xenografts analyzed is indicated in the images. Statistical results: ns > 0.05, * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, **** p ≤ 0.0001.

Article Snippet: Triple negative breast cancer cell lines Hs578T and MDA-MB-468, originally from American Type Culture Collection, were authenticated through short tandem repeat profiling karyotyping isoenzyme analysis.

Techniques: Labeling, Injection, Control, Immunopeptidomics, Staining, Confocal Microscopy

Journal: Cancers

Article Title: Zebrafish Xenografts Unveil Sensitivity to Olaparib beyond BRCA Status

doi: 10.3390/cancers12071769

Figure Lengend Snippet: Olaparib and IR can have modulating effects on the tumor vascular network. All cell lines were fluorescently labeled with CM-DiI (in red) and injected in the perivitelline space (PVS) of 2dpf Tg(fli1:eGFP) zebrafish embryos. At 24 hpi, zebrafish xenografts were screened and randomly distributed into the different experimental conditions: control, olaparib, IR, IR + olaparib. Xenografts were treated for 4 consecutive days, fixed at 5dpi and imaged by confocal microscopy for analysis of vessel density ( A – T ). The quantification of vessel density is represented by percentage of tumor area occupied by vessels (in green) ( U ). To analyze vessel functionality, TNBC cell line Hs578T was fluorescently labeled with CellTracker™ Deep Red Dye (in red-false color) and injected in the PVS of 2dpf Tg(gata1:RFP;fli1:eGFP). Hs578T xenografts were screened and randomly distributed in the different experimental conditions: control, olaparib, IR and IR + olaparib. Representative images of 5dpi Hs578T xenografts ( V , V’ ). The presence of erythrocytes (in red) inside the tumor-associated vessels (in green) was scored as: absence of erythrocytes = no perfusion ( W , W’ ) or presence of erythrocytes = with perfusion ( X , X’ ) and quantified ( Z ). All images are anterior to the left, posterior to right, dorsal up, and ventral down (as depicted in the scheme on top left). The dashed lines delineate the tumor area. White arrowheads indicate erythrocytes inside the vasculature. Scale bar: 50 µm. Results are from three independent experiments and expressed as mean ± SEM, each dot represents one xenograft. The total number of xenografts analyzed is indicated in the images. Statistical results: ns > 0.05, * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, **** p ≤ 0.0001.

Article Snippet: Triple negative breast cancer cell lines Hs578T and MDA-MB-468, originally from American Type Culture Collection, were authenticated through short tandem repeat profiling karyotyping isoenzyme analysis.

Techniques: Labeling, Injection, Control, Confocal Microscopy

Journal: Cancers

Article Title: Zebrafish Xenografts Unveil Sensitivity to Olaparib beyond BRCA Status

doi: 10.3390/cancers12071769

Figure Lengend Snippet: Metastatic potential of TNBC cell lines is modulated by olaparib and IR. TNBC (Hs578T BRCAwt , MDA-MB-469 BRCAwt and Hcc1937 BRCA1mut ) xenografts were generated as previously described and at 24 hpi, were screened and randomly distributed into the different experimental conditions: control, olaparib, IR, IR + olaparib. At 5dpi and 4 days post-treatment, xenografts were fixed to analyze the presence of micrometastasis in the caudal hematopoietic tissue (CHT) ( A ). Representative image of a micrometastasis in the CHT is labelled with anti-human-HLA–MHC-class (in red) for human cell identification. The dashed white line represents the micrometastasis area, scale bar: 50 µm ( A ). The metastatic potential was quantified as the percentage of xenografts that present micrometastasis in the CHT at 5dpi ( B ). Results are from three independent experiments and expressed as mean ± SEM. The total number of xenografts analyzed is indicated below the chart. Statistical results: ns > 0.05, * p ≤ 0.05.

Article Snippet: Triple negative breast cancer cell lines Hs578T and MDA-MB-468, originally from American Type Culture Collection, were authenticated through short tandem repeat profiling karyotyping isoenzyme analysis.

Techniques: Generated, Control